Linkage mapping and identification of QTL affecting deoxynivalenol (DON) content (Fusarium resistance) in oats (Avena sativa L.)

Mycotoxins caused by Fusarium spp. is a major concern on food and feed safety in oats, although Fusarium head blight (FHB) is often less apparent than in other small grain cereals. Breeding resistant cultivars is an economic and environment-friendly way to reduce toxin content, either by the identification of resistance QTL or phenotypic evaluation. Both are little explored in oats. A recombinant-inbred line population, Hurdal × Z595-7 (HZ595, with 184 lines), was used for QTL mapping and was phenotyped for 3 years. Spawn inoculation was applied and deoxynivalenol (DON) content, FHB severity, days to heading and maturity (DH and DM), and plant height (PH) were measured. The population was genotyped with DArTs, AFLPs, SSRs and selected SNPs, and a linkage map of 1,132 cM was constructed, covering all 21 oat chromosomes. A QTL for DON on chromosome 17A/7C, tentatively designated as Qdon.umb-17A/7C, was detected in all experiments using composite interval mapping, with phenotypic effects of 12.2–26.6 %. In addition, QTL for DON were also found on chromosomes 5C, 9D, 13A, 14D and unknown_3, while a QTL for FHB was found on 11A. Several of the DON/FHB QTL coincided with those for DH, DM and/or PH. A half-sib population of HZ595, Hurdal × Z615-4 (HZ615, with 91 lines), was phenotyped in 2011 for validation of QTL found in HZ595, and Qdon.umb-17A/7C was again localized with a phenotypic effect of 12.4 %. Three SNPs closely linked to Qdon.umb-17A/7C were identified in both populations, and one each for QTL on 5C, 11A and 13A were identified in HZ595. These SNPs, together with those yet to be identified, could be useful in marker-assisted selection to pyramiding resistance QTL.     

Anther extrusion and plant height are associated with Type I resistance to Fusarium head blight in bread wheat line ‘Shanghai-3/Catbird’

Fusarium head blight (FHB) is a destructive wheat disease of global importance. Resistance breeding depends heavily on the Fhb1 gene. The CIMMYT line Shanghai-3/Catbird (SHA3/CBRD) is a promising source without this gene. A recombinant inbred line (RIL) population from the cross of SHA3/CBRD with the German spring wheat cv. Naxos was evaluated for FHB resistance and related traits in field trials using spray and spawn inoculation in Norway and point inoculation in China. After spray and spawn inoculation, FHB severities were negatively correlated with both anther extrusion (AE) and plant height (PH). The QTL analysis showed that the Rht-B1b dwarfing allele co-localized with a QTL for low AE and increased susceptibility after spawn and spray inoculation. In general, SHA3/CBRD contributed most of the favorable alleles for resistance to severity after spray and spawn inoculation, while Naxos contributed more favorable alleles for reduction in FDK and DON content and resistance to severity after point inoculation. SHA3/CBRD contributed a major resistance QTL close to the centromere on 2DLc affecting FHB severity and DON after all inoculation methods. This QTL was also associated with AE and PH, with high AE and tall alleles contributed by SHA3/CBRD. Several QTL for AE and PH were detected, and low AE or reduced PH was always associated with increased susceptibility after spawn and spray inoculation. Most of the other minor FHB resistance QTL from SHA3/CBRD were associated with AE or PH, while the QTL from Naxos were mostly not. After point inoculation, no other QTL for FHB traits was associated with AE or PH, except the 2DLc QTL which was common across all inoculation methods. Marker-assisted selection based on the 2DLc QTL from SHA3/CBRD combined with phenotypic selection for AE is recommended for resistance breeding based on this valuable source of resistance.     

Folding and stability of outer membrane protein A (OmpA) from Escherichia coli in an amphipathic polymer, amphipol A8-35

Amphipols are a class of amphipathic polymers designed to maintain membrane proteins in aqueous solutions in the absence of detergents. Denatured β-barrel membrane proteins, like outer membrane proteins OmpA from Escherichia coli and FomA from Fusobacterium nucleatum, can be folded by dilution of the denaturant urea in the presence of amphipol A8-35. Here, the folding kinetics and stability of OmpA in A8-35 have been investigated. Folding is well described by two parallel first-order processes, whose half-times, ~5 and ~70 min, respectively, are independent of A8-35 concentration. The faster process contributed ~55–64 % to OmpA folding. Folding into A8-35 was faster than into dioleoylphosphatidylcholine bilayers and complete at ratios as low as ~0.17 g/g A8-35/OmpA, corresponding to ~1–2 A8-35 molecules per OmpA. Activation energies were determined from the temperature dependence of folding kinetics, monitored both by electrophoresis, which reports on the formation of stable OmpA tertiary structure, and by fluorescence spectroscopy, which reflects changes in the environment of tryptophan side chains. The two methods yielded consistent estimates, namely ~5–9 kJ/mol for the fast process and ~29–37 kJ/mol for the slow one, which is lower than is observed for OmpA folding into dioleoylphosphatidylcholine bilayers. Folding and unfolding titrations with urea demonstrated that OmpA folding into A8-35 is reversible and that amphipol-refolded OmpA is thermodynamically stable at room temperature. Comparison of activation energies for folding and unfolding in A8-35 versus detergent indicates that stabilization of A8-35-trapped OmpA against denaturation by urea is a kinetic, not a thermodynamic phenomenon.     

Effect of different C/N ratios on carotenoid and lipid production by Rhodotorula glutinis

Due to the increasing demand for sustainable biofuels, microbial oils as feedstock for the transesterification into biodiesel have gained scientific and commercial interest. Also, microbial carotenoids have a considerable market potential as natural colorants. The carbon to nitrogen (C/N) ratio of the respective cultivation media is one of the most important parameters that influence the production of microbial lipids and carotenoids. Thus, in the present experiment, the influence of different C/N ratios, initial glucose loadings, and ammonium concentrations of the cultivation medium on microbial cell growth and lipid and carotenoid production by the oleaginous red yeast Rhodotorula glutinis has been assessed. As a general trend, both lipid and carotenoid production increased at high C/N ratios. It was shown that not only the final C/N ratio but also the respectively applied initial carbon and nitrogen contents influenced the observed parameters. The lipid yield was not affected by different ammonium contents, while the carotenoid production significantly decreased both at low and high levels of ammonium supply. A glucose-based increase from C/N 70 to 120 did not lead to an increased lipid production, while carotenoid synthesis was positively affected. Generally, it can be asserted that lipid and carotenoid synthesis are stimulated at higher C/N ratios.     

Enzyme-catalyzed protein crosslinking

The process of protein crosslinking comprises the chemical, enzymatic, or chemoenzymatic formation of new covalent bonds between polypeptides. This allows (1) the site-directed coupling of proteins with distinct properties and (2) the de novo assembly of polymeric protein networks. Transferases, hydrolases, and oxidoreductases can be employed as catalysts for the synthesis of crosslinked proteins, thereby complementing chemical crosslinking strategies. Here, we review enzymatic approaches that are used for protein crosslinking at the industrial level or have shown promising potential in investigations on the lab-scale. We illustrate the underlying mechanisms of crosslink formation and point out the roles of the enzymes in their natural environments. Additionally, we discuss advantages and drawbacks of the enzyme-based crosslinking strategies and their potential for different applications.     

Changes in fatty acid composition,cholesterol and fat‐soluble vitamins during development of eggs and larvae in shabbout (Barbus grypus,Heckel 1843)

Changes in fatty acid composition, cholesterol and fat‐soluble vitamins were studied during development (fertilized eggs, yolk‐sac larvae, and after yolk resorption of shabbout, Barbus grypus). Significant differences were found in the monounsaturated fatty acids (MUFAs), polyunsaturated fatty acids (PUFA), PUFA/saturated fatty acids (SFA), ∑n‐3 and eicosapentaenoic acid (EPA)/docosahexaenoic acid (DHA) ratios between eggs and larvae (P < 0.05). Significant differences were also observed in the C14:0, C16:1n‐7, C18:1n‐9, C18:3n‐6, C20:0, C20:4n‐6, C24:0, C24:1, C22:6n‐3 fatty acids between eggs and larvae after yolk‐sac resorption (P < 0.05). Vitamin α‐ Tocopherol and retinol content increased during embryogenesis, but changes were insignificant in retinol acetate, δ‐Tocopherol, K1, K2 and cholesterol content between eggs and larvae after yolk resorption (P > 0.05).     

Is the Soft Song of the Brownish‐Flanked Bush Warbler an Aggressive Signal?

Soft songs have been detected in many songbirds, but in most species, research on soft songs has lagged behind studies of broadcast songs. In this study, we describe the acoustic features of a soft song in the brownish‐flanked bush warbler Cettia fortipes. Compared with the broadcast song, the warbler's soft song was characterized by a lower minimum frequency and longer duration, and it had a higher proportion of rapid frequency modulation notes. Using playback experiments, some in combination with mounted specimens, we found different responses to soft and broadcast songs, and we found that soft song can predict aggressive escalation (attack). We conclude that the soft song is an aggressive signal in this species.     

Trapping of European buprestid beetles in oak forests using visual and olfactory cues

Trapping approaches developed for the emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), were adapted for trapping several European oak buprestid species. These approaches included the use of natural leaf surfaces as well as green and purple plastic in sticky trap designs. Plastic surfaces were incorporated into novel ‘branch‐trap’ designs that each presented two 5 × 9‐cm² rectangular surfaces on a cardboard structure wrapped around the leaves of a branch. We used visual adult Agrilus decoys in an attempt to evoke male mating approaches toward the traps. Our first experiment compared the attractiveness of visual characteristics of the surfaces of branch‐traps. The second looked at the effect on trap captures of adding semiochemical lures, including manuka oil, (Z)‐3‐hexen‐1‐ol, and (Z)‐9‐tricosene. In total, 1 962 buprestid specimens including 14 species from the genus Agrilus were caught on 178 traps in a 22‐day time‐span. Overall, the green plastic‐covered branch‐traps significantly out‐performed the other trap designs. We further found that the presence of an EAB visual decoy placed on the trap surface often increased captures on these green traps, but this effect was stronger for certain Agrilus species than for others. The visual decoy was particularly important for the most serious pest detected, Agrilus biguttatus Fabricius, which was captured 13 times on traps with decoys, but only once without a decoy. There were some small but significant effects of odor treatment on the capture of buprestids of two common species, Agrilus angustulus Illiger and Agrilus sulcicollis Lacordaire. There were also 141 Elateridae specimens on these traps, which were not influenced by trap type or decoys. The results suggest that small branch‐traps of this nature can provide a useful new tool for monitoring of buprestids, which have the potential to be further optimized with respect to visual and olfactory cues.